Multispecies

SUPPLEMENTARY DATA
Author

Amélie Lehuen, Rose-Marie Oulhen, Zhengquan Zhou, Jaco de Smit, Lennart van Ijzerloo, Francesco Cozzoli, Tjeerd Bouma, Francis Orvain

Published

May 23, 2024

1 Introduction

NB: Graphs can be observed in full screen by a right click + “Open image in new tab”

2 Materials and Methods

All data processing was conducted in R version 4.4.0 (2024-04-24 ucrt) and Matlab 2021a. Significance levels are p < .0001 with “****”, p < .001 with “***”, p < .01 with “**”, p < .05 with “*”.

2.1 Biological models

SuppFig 2.A: Fauna models used in their sediment.
SuppFig 2.B: Fauna models used in their sediment.

Metabolic rate calculation

A model for Mass Specific Respiration rate (MSR) of aquatic invertebrates was developed by Brey (Brey 2010), providing a spreadsheet tool that implement an Artificial Neural Network (ANN). It requires as arguments : (1) individual body mass in J, (2) temperature, (3) depth in water, (4) taxonomic definition, (5) mobility mode (sessile, crawler, facultative or permanent swimmer/floater), (6) alimentation mode (carnivorous versus other modes), (7) vision type (yes or no defined as possession of image-forming eyes sensu (Seibel and Drazen 2007), i.e. an optical sense better than just light/dark separation), and (8) the starved state of the animal (yes or no). MSR calculation was made using the average energy density 21.4469 J.mgAFDW^{-1} (Brey et al. 2010), a depth of 1 m for intertidal species, and by default not starved. A conversion was done for the MSR from [J/J/day] to [mW.ind^{-1}]. Based on the theoretical frame of Brown and Allen ((Allen, Gillooly, and Brown 2005, @brown2004)), the individual metabolic rate I was expressed as function of the individual body size and the temperature. The equation was simplified to adjust the parameters r0, b and Ek for each species to the MSR results based on Brey ANN (Equation 2.A, Table 2.A).

I = r0.(IndivBodySize)^b . e^{\frac{-Ek}{(T+273,15)}} \tag{2.A}

Itot = I . Density \tag{2.B}

With the exponential part the factor Ek [K] is the ratio of E, activation energy over k Boltzmann’s constant (8.62×10−5 eV.K−1); r0 is the scaling constant; b is the scaling exponent; T is temperature [°C], IndivBodySize is the individual body size in Ash Free Dry Weigth [gAFDW.ind^{-1}], to get I [mW.ind{-1}]. Density is the spatial density of individuals in flume (based on sample surface) [ind.m^{-2}], thus Itot [mW.m^{-2}].

SuppTab 2.A: Species Brey Specific Metabolic Rate inputs and equation parameters results for Equation 2.A.
Species_Name AphiaId TaxaBrey Mobility Trophic Vision b r0 Ek
Hediste diversicolor 152302 Annelida Crawl Deposit feeder 0 0.7733019 3.386977e+09 6197.232
Scrobicularia plana 141424 Mollusca 1 Sessile Mixt Filter feeder 0 0.7752776 5.559549e+13 8986.158

Bioturbation processes

As far as the consequences on sediment erodibility are concerned, the influence of bioturbation should rather be divided into two main categories Figure 2.C: 1) whether the effect is chronic and generates a biogenic layer under standard hydrodynamic conditions (Mariotti and Fagherazzi 2012) or 2) whether it has an impact on the constituent layers of the sediment and is expressed in the case of high hydrodynamic events (mass erosion, roughly above 1Pa bed shear stress). Second, these effects can be characterized according to the type of consequence: either stabilizing by reinforcing physical protection and cohesion between particles or destabilizing by increasing surface roughness or modifying the rheology of sediments facilitating erosion when tidal currents or wave-induced turbulence are present Willows, Widdows, and Wood (1998). Several processes are possible in each case, for chronic effects, two kinds of processes were identified:

  1. Passive processes relative to hydrodynamics modifications: 1a) Allogenic skimming flow operates when a high density of biogenic structures creates locally reduced hydrodynamic conditions, favouring the deposition of fine particles and the consolidation of sediments; 1b) Autogenic roughness causes a modification of hydrodynamic conditions on a very local scale by increasing the bed shear stress and the flow regime.

  2. Active processes: 2a) Biodeposition is carried out by suspension feeders, which capture fine particles in suspension and agglomerate them into pseudo-faeces or faeces, leading to a muddification of the sedimentary bed. When combined with the biodiffusion effect, it increases particle mixing and sediment stability, while also promoting the development of MPB biofilms; 2b) Bioresuspension occurs when individuals produce tracks in relation to displacement or feeding behaviours (producing faecal pellets and/or pseudo- faeces) on the sediment surface, therefore generating a biogenic fluff layer. The amount of sediment reworked by this process may be limited by a high density or by a sufficient duration that will lead the bioturbating fauna to rework sediments that have already been reworked.

For the event-driven cases, two main processes were identified: 1) Biostabilization is a stabilisation against mass erosion by sedentary species which, by settling in galleries or tubes, compact the sediment and bind the particles together with their mucus; 2) Bed destabilization causes anticipated mass erosion of the sediment assimilated to a massive loss of the surficial part of sediment at a centimetre scale, and occurs when endogenous species are mobile in the sediment (biodiffusors or regenerators).

SuppFig 2.C: The different types of bioturbation processes according to Kristensen et al. (2012) plus processes linked to sediment erodibility. Processes with green stars are stabilizing, with red stars are destabilizing.
SuppTab 2.B: Bibliographical references for the classification of the effects of bioturbation on sediment erodibility for the six model species.

2.2 Sediment and animal collection

The Schelde estuary, a macrotidal coastal estuary, situated between The Netherlands and Belgium, is split in two main parts, named Westerschelde (south part) and Oosterschelde (north part). Due to anthropological transformations, the Oosterschelde is no longer fed by Schelde freshwater (Figure 2.D).

SuppFig 2.D: Maps of sampling location for each species and sediment.

A muddy and a sandy sediment were collected in the Westerschelde (respectively mud in Groot Buitenschoor; sand in Rilland), wet-sieved at 5mm and stored 48h in a freezer, then wet-sieved at 1mm, to remove fauna and larger debris. Each sediment granulometry was characterized with a Mastersizer 2000 (Malvern Instruments Ltd., Malvern, UK). A 50%-50% vol mix was made, an let settle for a couple of weeks to reduce the water content. The water content, density, granulometry and organic matter were monitored all along the experiment. The granulometry of the stockpile is analysed once a week. The organic matter (OM) was obtained by loss on ignition: samples were placed in an oven for 72 h to remove the water and then placed in an oven at 550°C for 6 h to burn the OM (results).

SuppFig 2.E: Main sizes particules sediment granulometry: in brown stars, mud and sand prior mixing, in grey circles sediment from fauna sampling sites, in colored circles, experimental sediment with its evolution during experiment.
SuppTab 2.C: Granulometry table
Field Source Stock
Parameter 2022-07-04 2022-07-15 2022-07-26 2022-06-03 2022-06-13 2022-06-17 2022-06-24 2022-07-01 2022-07-08 2022-07-13 2022-07-25 2022-07-29
MACBA-SCRPL CERED CORVO HEDDI PERUL Mud Sand Stock1 Stock2 Stock3 Stock4 Stock5 Stock6 Stock7 Stock8 Stock9
10% limit (10% < ...μm) 3.88 175.63 5.91 6.67 3.73 3.23 95.67 6.96 4.94 4.82 5.16 5.54 5.21 5.75 6.32 6.44
Median grainsize D50 in μm 28.00 287.05 51.51 52.99 25.33 20.69 158.83 82.25 53.14 45.99 68.97 62.91 58.58 70.82 81.31 87.92
Median grainsize D50 in PHI 5.16 1.80 4.28 4.24 5.30 5.60 2.65 3.60 NA NA 3.86 3.99 4.09 3.82 3.62 3.51
Modus grainsize in μm 101.38 288.64 71.38 81.78 24.38 26.22 161.69 161.56 152.04 153.24 154.37 155.68 155.49 161.60 160.72 159.17
90% limit (90% < ...μm) 137.28 465.87 119.97 131.43 121.21 77.69 252.42 235.56 203.85 204.16 221.03 222.61 220.46 232.34 233.28 233.25
Coarse sand fraction PHI 0-1, 500-1000 μm 0.00 6.74 0.11 0.21 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00
Medium sand fraction PHI 1-2, 250-500 μm 0.66 56.89 0.53 0.32 0.81 0.83 10.53 8.07 4.02 4.19 6.28 6.61 6.32 7.72 7.77 7.71
Fine sand fraction PHI 2-3, 125-250 μm 11.90 35.70 8.08 11.17 8.56 3.13 62.81 29.04 26.71 25.77 27.48 26.10 26.18 27.73 29.05 30.27
Very Fine sand fraction PHI 3-4, 62.5-125 μm 19.75 0.68 31.89 31.52 16.10 10.44 23.15 17.54 17.23 16.33 17.53 17.39 16.74 16.18 17.49 18.01
Silt fraction <63 μm 67.89 0.00 59.80 57.13 74.73 85.79 3.53 45.46 52.04 53.71 48.80 50.02 50.87 48.48 45.80 44.11
SuppFig 2.F: Water content rate evolution during experiment and each temperature condition regression.
SuppFig 2.G: Water content rate versus temperature condition. The T high condition had significantly lower water content than the two other conditions.

Species were collected either in Oosterschelde or Westerschelde; C. edule in Oesterdam at first sampling then in Den Inkel; H. diversicolor in Haven Rattekaai; C. volutator in Haventje Ellewoutsdijk; S. plana and M. balthica in Den Inkel; P. ulvae in Nolleweg. Individuals of each species were sorted to create batches of size classes, and a sub-sample of each class were used to measure length, fresh weight, dry weight, Ash Free Dry Weight and define conversion coefficients that were used to create sample populations. The rest of the individuals were placed in the climatized mesocosm, an acclimation period of 2 weeks was respected before experiments.

Biometry done on each species and relation between all dimensions. Length: Length of shell [mm]; FW_shell: fresh weight with shell [g], FW: fresh weight of flesh [g], DW: Dry weight of flesh [g] (48h at 60°C), AFDW: ash free dry weight [g] (5h at 550°C).

Biometry done on each species and relation between all dimensions. Length: Length of shell [mm]; FW_shell: fresh weight with shell [g], FW: fresh weight of flesh [g], DW: Dry weight of flesh [g] (48h at 60°C), AFDW: ash free dry weight [g] (5h at 550°C).

2.3 Experimental design

SuppFig 2.H: Experimental factors combinations.
SuppFig 2.I: Mesocosms tidal rhythm for each duo. The blue arrow represents the respiration measure and the installation in the pot, the red arrow the bioturbation phase.
SuppFig 2.J: Sample biological levels in number of individuals on cores

2.4 Experimental measurement

SuppTab 2.D: Data set summary
Characteristic Control Small S. plana & H. diversicolor Medium S. plana & H. diversicolor Big S. plana & H. diversicolor
T. medium, N = 61 T. low, N = 31 T. high, N = 31 T. medium, N = 81 T. low, N = 81 T. high, N = 81 T. medium, N = 81 T. low, N = 81 T. high, N = 91 T. medium, N = 81 T. low, N = 81 T. high, N = 91
Experimental design
Ambiant temperature (°C) 17.85+/-0.00 17.50+/-0.00 24.65+/-0.00 18.38+/-2.79 16.55+/-0.41 20.38+/-3.99 18.38+/-2.79 16.55+/-0.41 20.57+/-3.78 18.38+/-2.79 16.55+/-0.41 20.57+/-3.78
Mesocosm temperature (°C) 13.28+/-0.01 9.32+/-0.00 18.20+/-0.00 13.28+/-0.23 9.33+/-0.62 18.20+/-1.15 13.28+/-0.23 9.32+/-0.62 18.19+/-1.07 13.26+/-0.22 9.32+/-0.62 18.19+/-1.07
Metabolic rate in mesocosm (mW/m²) 0.00-0.00 0.00-0.00 0.00-0.00 9.57-19.72 5.88-10.50 15.31-32.25 17.48-29.99 12.05-19.47 22.03-58.14 25.61-46.29 18.43-31.05 42.82-81.93
Species 1 Metabolic rate in mesocosm (mW/m²) 0.00-0.00 0.00-0.00 0.00-0.00 0.00-19.72 0.00-10.50 0.00-32.25 0.00-29.99 0.00-16.58 0.00-58.14 0.00-46.29 0.00-25.82 0.00-81.93
Species 2 Metabolic rate in mesocosm (mW/m²) 0.00-0.00 0.00-0.00 0.00-0.00 0.00-13.84 0.00-8.85 0.00-24.41 0.00-20.44 0.00-19.47 0.00-43.50 0.00-35.33 0.00-31.05 0.00-64.03
Species 1 density in sample (ind/m²) 0.00-0.00 0.00-0.00 0.00-0.00 0.00-160.00 0.00-160.00 0.00-160.00 0.00-160.00 0.00-160.00 0.00-160.00 0.00-160.00 0.00-160.00 0.00-160.00
Species 2 density in sample (ind/m²) 0.00-0.00 0.00-0.00 0.00-0.00 0.00-320.00 0.00-320.00 0.00-320.00 0.00-320.00 0.00-320.00 0.00-320.00 0.00-320.00 0.00-320.00 0.00-320.00
Species 1 biomass in sample (gAFDW/m²) 0.00-0.00 0.00-0.00 0.00-0.00 0.00-7.56 0.00-6.17 0.00-5.84 0.00-12.99 0.00-11.13 0.00-12.48 0.00-22.87 0.00-19.94 0.00-19.53
Species 2 biomass in sample (gAFDW/m²) 0.00-0.00 0.00-0.00 0.00-0.00 0.00-3.61 0.00-3.17 0.00-4.09 0.00-5.98 0.00-8.71 0.00-8.63 0.00-12.66 0.00-15.93 0.00-14.23
Metabolic rate based on respiration meas (mW/m²) 0.00+/-0.00 0.00+/-0.00 0.00+/-0.00 13.20+/-8.76 12.25+/-9.76 23.74+/-9.75 30.61+/-5.64 17.38+/-6.28 41.85+/-15.14 34.59+/-25.24 41.76+/-12.44 65.71+/-25.64
Results
Bed Shear Stress (Pa) 7.31+/-0.51 3.32+/-1.26 3.13+/-0.49 4.43+/-2.50 2.49+/-1.36 4.52+/-1.20 2.51+/-1.32 2.69+/-1.57 3.81+/-1.26 2.55+/-1.26 3.06+/-1.65 3.28+/-1.32
1 Mean+/-SD; Minimum-Maximum
SuppFig 2.K: Sample composition for each duo in metabolic rates (MSR calculated with mesocosm T°).

3 Results

3.1 Respiration measurements

SuppFig 3.A: All respirometry results for OsCar runs
SuppFig 3.B: All respirometry results for OsCar runs
SuppTab 3.A: Respiration data summary
Characteristic Small S. plana & H. diversicolor Medium S. plana & H. diversicolor Big S. plana & H. diversicolor
T. low, N = 8 T. medium, N = 10 T. high, N = 9 T. low, N = 9 T. medium, N = 10 T. high, N = 10 T. low, N = 9 T. medium, N = 10 T. high, N = 10
Temperature mesocosm (°C)








    Mean (SD) 9.33 (0.62) 13.28 (0.20) 18.20 (1.07) 9.32 (0.58) 13.28 (0.20) 18.19 (1.01) 9.32 (0.58) 13.26 (0.19) 18.19 (1.01)
    Range 8.87, 10.83 13.07, 13.70 17.23, 20.08 8.87, 10.83 13.07, 13.70 17.23, 20.08 8.87, 10.83 13.07, 13.70 17.23, 20.08
Respiration Rate (µmolO2/s-1)








    Mean (SD) 0.00 (0.00) 0.00 (0.00) 0.00 (0.00) 0.00 (0.00) 0.00 (0.00) 0.00 (0.00) 0.00 (0.00) 0.00 (0.00) 0.00 (0.00)
    Range 0.00, 0.00 0.00, 0.00 0.00, 0.00 0.00, 0.00 0.00, 0.00 0.00, 0.00 0.00, 0.00 0.00, 0.00 0.00, 0.01
Itot based on respiration meas (mW/m²)








    Mean (SD) 12.25 (9.76) 10.56 (9.52) 21.10 (12.07) 15.45 (8.25) 24.49 (13.83) 37.67 (19.47) 37.12 (18.14) 27.67 (26.61) 59.13 (31.88)
    Range 0.00, 29.59 0.00, 26.95 0.00, 37.36 0.00, 24.60 0.00, 36.40 0.00, 67.75 0.00, 61.20 0.00, 85.51 0.00, 97.42
Respiration Rate (µmolO2/gAFDW-1)








    Mean (SD) 0.21 (0.14) 0.19 (0.15) 0.29 (0.11) 0.12 (0.06) 0.21 (0.05) 0.27 (0.11) 0.16 (0.07) 0.15 (0.14) 0.24 (0.10)
    Range 0.05, 0.50 0.02, 0.47 0.14, 0.48 0.02, 0.20 0.14, 0.29 0.07, 0.44 0.08, 0.28 0.00, 0.43 0.08, 0.38
    Unknown 1 2 1 1 2 1 1 2 1
SuppFig 3.C: ANCOVA results
SuppTab 3.B: ANCOVA results
Respiration_rate_mWm2..y_tf ~ Itot_meso_mWm2_Respi..x_tf + Mumo_txt + Cond_Fact_txt
Characteristic Beta1 SE2 p-value
(Intercept) 0.8877*** 1.535 × 10−4 2.350 × 10−187
Itot at mesocosm temperature - transf (mW/m²) 0.0001*** 1.331 × 10−5 8.377 × 10−13
Mumo_txt NA***
9.506 × 10−5
    S. plana -0.0004*** 8.219 × 10−5 1.600 × 10−5
    S. plana & (H. diversicolor) 0.0000 8.011 × 10−5 0.833
    (S. plana) & H. diversicolor 0.0002** 7.982 × 10−5 0.004
    H. diversicolor 0.0001 8.000 × 10−5 0.163
Cond_Fact_txt NA
0.208
    T. low 0.0001 7.666 × 10−5 0.078
    T. medium 0.0000 6.556 × 10−5 0.515
    T. high -0.0001 8.096 × 10−5 0.37
No. Obs. = 72; R² = 0.646; AIC = -920
Anova, F(2,65) = 1.61, p = 0.21, eta2[g] = 0.047
Respiration_rate_mWm2..y_tf ~ Itot_meso_mWm2_Respi..x_tf + Mumo_txt + Cond_Fact_txt post-hoc tests
Residuals are normal - Shapiro-Wilk: stat = 0.977, p-value = 0.213 - (α=0.1)
Residuals variances are homogeneic - Levene: stat = 0.987, p-value = 0.468 - (α=0.1)
Residuals are homoscedastic - Harrison-McCabe: statistic = 0.448, p-value = 0.246 (α=0.1)
There are 0 outliers in **
1 *p<0.05; **p<0.01; ***p<0.001
2 SE = Standard Error
SuppTab 3.C: Data set description and application conditions
Variable Control, N = 01 S. plana, N = 191 S. plana & (H. diversicolor), N = 181 (S. plana) & H. diversicolor, N = 181 H. diversicolor, N = 171 Overall, N = 721
Respiration_rate_mWm2 (dependent) NA (NA) 29 (22) 28 (12) 36 (22) 39 (28) 33 (22)
Itot at mesocosm temperature (mW/m²) NA (NA) 34 (21) 23 (11) 22 (12) 26 (16) 26 (16)
Temperature level





    T. low 0 6 5 6 6 23
    T. medium 0 6 6 6 5 23
    T. high 0 7 7 6 6 26
Respiration_rate_mWm2 normality test
Data are not normal - Shapiro-Wilk: stat=0.900, p-value=3.06e-05 (α=0.1).
Data transformation - Box-Cox λ = 0.340; λ2 = 1
Transformed data are normal - Shapiro-Wilk: stat = 0.984, p-value = 0.505 (α=0.1)
Itot_meso_mWm2_Respi normality test
Data are not normal - Shapiro-Wilk: stat=0.943, p-value=0.00279 (α=0.1).
Data transformation - Box-Cox λ = 0.010; λ2 = 1
Transformed data are normal - Shapiro-Wilk: stat = 0.991, p-value = 0.875 (α=0.1)
Treatments subgroup check
Variances are homogeneic - Levene: F value = 0.609, p-value = 0.814 - (α=0.1)
Independence of covariate: Mumo_txt; Cond_Fact_txt have a significant relation with covariate
Regression slopes homogeneity is verified
Respiration_rate_mWm2..y_tf ~ Itot_meso_mWm2_Respi..x_tf + Mumo_txt + Cond_Fact_txt post-hoc tests
Residuals are normal - Shapiro-Wilk: stat = 0.977, p-value = 0.213 - (α=0.1)
Residuals variances are homogeneic - Levene: stat = 0.987, p-value = 0.468 - (α=0.1)
Residuals are homoscedastic - Harrison-McCabe: statistic = 0.448, p-value = 0.252 (α=0.1)
There are 0 outliers in **
1 Mean (SD); n

3.2 Erodibility analysis

3.2.1 Erosion data treatment

3.2.2 Erosion measurement treatment

SuppFig 3.D: Sketch of erosion processes in the fluff layer erosion (green line), mass erosion (solid red line) and the parameters used. The dashed black line represents the addition of the two processes, the SSC measured in the experiment. The straight dashed black line represents the linear regression of the mass erosion steps that determined the critical mass erosion threshold ([Pa]).
SuppTab 3.D: Erosion data summary
Characteristic 0, N = 9311 1, N = 4441 2, N = 4681
Step number 6 (3, 10) 16 (12, 19) 16 (13, 19)
Sec_Palier 393+/-318 859+/-255 910+/-254
Bed shear stress (Pa)


    Mean+/-SD 1.86+/-1.87 5.16+/-1.95 5.31+/-1.85
    Range 0.06 - 8.81 0.64 - 9.95 1.10 - 10.15
Shear velocity (cm.s-1)


    Mean+/-SD 3.77+/-1.97 6.95+/-1.43 7.08+/-1.29
    Range 0.74 - 9.28 2.51 - 9.86 3.28 - 9.95
1 Median (IQR); Mean+/-SD
SuppFig 3.E: Controls data ANOVA results versus temperature for A) Qfluff and B) BSS mass threshold

3.2.3 Mass erosion threshold

SuppFig 3.F: Critical bed shear stress (BSSmass [Pa]) vs the different metabolic rate evaluation [mW.m^{-2}]: the measured respiration rate (A), the Itot with the mesocosm temperature (B) and the Itot with the flume temperature (C) and their regression line. Note that the scales are Box-Cox transformed, and the models were made on transformed data.
SuppFig 3.G: ANCOVA results
SuppTab 3.E: ANCOVA results
BSS_Pa_Crit_Mass_Ssc..y_tf ~ S1_Itot_meso_mWm2_Ero + S2_Itot_meso_mWm2_Ero + Mumo_txt
Characteristic Beta1 SE2 p-value
(Intercept) 1.3487*** 0.079 8.151 × 10−28
S. plana Itot (mW/m²) -0.0025 0.004 0.532
H. diversicolor Itot (mW/m²) 0.0132* 0.006 0.038
Duo NA*
0.018
    Control 0.3743** 0.119 0.002
    S. plana 0.1284 0.139 0.358
    S. plana & (H. diversicolor) -0.0131 0.084 0.876
    (S. plana) & H. diversicolor -0.1238 0.089 0.167
    H. diversicolor -0.3659* 0.141 0.029
No. Obs. = 84; R² = 0.148; AIC = 85.3
Anova, F(4,77) = 3.18, p = 0.018, eta2[g] = 0.142
BSS_Pa_Crit_Mass_Ssc..y_tf ~ S1_Itot_meso_mWm2_Ero + S2_Itot_meso_mWm2_Ero + Mumo_txt post-hoc tests
Residuals are not normal - Shapiro-Wilk: stat = 0.965, p-value = 0.0208 - (α=0.1)
Residuals variances are homogeneic - Levene: stat = 1.864, p-value = 0.125 - (α=0.1)
Residuals are homoscedastic - Harrison-McCabe: statistic = 0.554, p-value = 0.741 (α=0.1)
There are 0 outliers in **
1 *p<0.05; **p<0.01; ***p<0.001
2 SE = Standard Error
SuppFig 3.H: BSS versus metabolic rate of the two species in same sample
SuppTab 3.F: BSS vs metabolic rates of each species of duo regression coefficients
Mumo_txt Intercept S1 S2 gcd.r.squared p_signif Nb obs
BSS_Pa_Crit_Mass_Ssc All 1.46e+00 [1.31e+00, 1.61e+00] -2.16e-03 [-7.78e-03, 3.47e-03] -1.80e-03 [-9.95e-03, 6.34e-03] 0.008 84
Control 1.72e+00 [1.47e+00, 1.98e+00] NA [NA, NA] NA [NA, NA] 0.000 - 11
S. plana 1.42e+00 [9.74e-01, 1.86e+00] -7.65e-04 [-1.20e-02, 1.04e-02] NA [NA, NA] 0.001 19
S. plana & (H. diversicolor) 1.50e+00 [1.08e+00, 1.91e+00] -2.11e-04 [-3.71e-02, 3.66e-02] -1.73e-02 [-9.03e-02, 5.57e-02] 0.037 19
(S. plana) & H. diversicolor 1.33e+00 [9.93e-01, 1.66e+00] -1.09e-02 [-1.06e-01, 8.40e-02] 1.07e-02 [-4.05e-02, 6.18e-02] 0.024 18
H. diversicolor 8.45e-01 [4.12e-01, 1.28e+00] NA [NA, NA] 1.88e-02 [3.01e-03, 3.46e-02] 0.300 * 17
SuppFig 3.I: ANCOVA results
SuppTab 3.G: ANCOVA results
BSS_Pa_Crit_Mass_Ssc..y_tf ~ Itot_meso_mWm2_Ero + Cond_Fact_txt
Characteristic Beta1 SE2 p-value
(Intercept) 1.55275*** 0.076 <0.001
Itot at mesocosm temperature (mW/m²) -0.00635* 0.003 0.030
Cond_Fact_txt NA**
0.005
    T. medium 0.03184 0.059 0.6
    T. low -0.20741** 0.063 0.002
    T. high 0.17558* 0.067 0.015
No. Obs. = 84; R² = 0.131; AIC = 81.0
Anova, F(2,80) = 5.7, p = 0.005, eta2[g] = 0.125
BSS_Pa_Crit_Mass_Ssc..y_tf ~ Itot_meso_mWm2_Ero + Cond_Fact_txt post-hoc tests
Residuals are not normal - Shapiro-Wilk: stat = 0.969, p-value = 0.0406 - (α=0.1)
Residuals variances are homogeneic - Levene: stat = 1.500, p-value = 0.229 - (α=0.1)
Residuals are homoscedastic - Harrison-McCabe: statistic = 0.566, p-value = 0.832 (α=0.1)
There are 0 outliers in **
1 *p<0.05; **p<0.01; ***p<0.001
2 SE = Standard Error
SuppFig 3.J: ANCOVA results
SuppTab 3.H: ANCOVA results
BSS_Pa_Crit_Mass_Ssc..y_tf ~ Itot_meso_mWm2_Ero + Mumo_txt
Characteristic Beta1 SE2 p-value
(Intercept) 1.38715*** 0.078 <0.001
Itot at mesocosm temperature (mW/m²) 0.00247 0.003 0.4
Duo NA
0.086
    Control 0.33586** 0.120 0.006
    S. plana -0.07961 0.092 0.4
    S. plana & (H. diversicolor) -0.05926 0.082 0.5
    (S. plana) & H. diversicolor -0.05556 0.083 0.5
    H. diversicolor -0.14143 0.086 0.3
No. Obs. = 84; R² = 0.105; AIC = 87.5
Anova, F(4,78) = 2.12, p = 0.086, eta2[g] = 0.098
BSS_Pa_Crit_Mass_Ssc..y_tf ~ Itot_meso_mWm2_Ero + Mumo_txt post-hoc tests
Residuals are not normal - Shapiro-Wilk: stat = 0.969, p-value = 0.0393 - (α=0.1)
Residuals variances are homogeneic - Levene: stat = 1.475, p-value = 0.218 - (α=0.1)
Residuals are homoscedastic - Harrison-McCabe: statistic = 0.538, p-value = 0.692 (α=0.1)
There are 0 outliers in **
1 *p<0.05; **p<0.01; ***p<0.001
2 SE = Standard Error
SuppFig 3.K: ANCOVA results
SuppTab 3.I: ANCOVA results
BSS_Pa_Crit_Mass_Ssc..y_tf ~ Itot_meso_mWm2_Ero + Mumo_txt + Cond_Fact_txt
Characteristic Beta1 SE2 p-value
(Intercept) 1.49480*** 0.089 <0.001
Itot at mesocosm temperature (mW/m²) -0.00292 0.004 0.4
Duo NA
0.4
    Control 0.22246 0.125 0.079
    S. plana -0.01174 0.094 >0.9
    S. plana & (H. diversicolor) -0.05681 0.079 0.5
    (S. plana) & H. diversicolor -0.04548 0.081 0.6
    H. diversicolor -0.10843 0.085 0.5
Cond_Fact_txt NA*
0.040
    T. medium 0.03165 0.059 0.6
    T. low -0.17269* 0.068 0.013
    T. high 0.14104 0.071 0.077
No. Obs. = 84; R² = 0.177; AIC = 84.4
Anova, F(2,76) = 3.35, p = 0.04, eta2[g] = 0.081
BSS_Pa_Crit_Mass_Ssc..y_tf ~ Itot_meso_mWm2_Ero + Mumo_txt + Cond_Fact_txt post-hoc tests
Residuals are not normal - Shapiro-Wilk: stat = 0.965, p-value = 0.0212 - (α=0.1)
Residuals variances are homogeneic - Levene: stat = 1.141, p-value = 0.34 - (α=0.1)
Residuals are homoscedastic - Harrison-McCabe: statistic = 0.519, p-value = 0.6 (α=0.1)
There are 0 outliers in **
1 *p<0.05; **p<0.01; ***p<0.001
2 SE = Standard Error
SuppTab 3.J: Data set description and application conditions
Variable Control, N = 111 S. plana, N = 191 S. plana & (H. diversicolor), N = 191 (S. plana) & H. diversicolor, N = 181 H. diversicolor, N = 171 Overall, N = 841
BSS_Pa_Crit_Mass_Ssc (dependent) 5.08 (2.24) 3.50 (2.10) 3.29 (1.58) 3.20 (1.19) 3.06 (1.66) 3.50 (1.82)
Itot at mesocosm temperature (mW/m²) (covariate) 0 (0) 34 (21) 22 (11) 22 (12) 25 (13) 22 (17)
Temperature level





    T. medium 5 6 6 6 6 29
    T. low 3 6 6 6 6 27
    T. high 3 7 7 6 5 28
BSS_Pa_Crit_Mass_Ssc normality test
Data are not normal - Shapiro-Wilk: stat=0.945, p-value=0.00129 (α=0.1).
Data transformation - Box-Cox λ = -0.010; λ2 = 1
Transformed data are normal - Shapiro-Wilk: stat = 0.983, p-value = 0.352 (α=0.1)
Treatments subgroup check
Variances are homogeneic - Levene: F value = 1.179, p-value = 0.311 - (α=0.1)
Independence of covariate: Mumo_txt; Cond_Fact_txt have a significant relation with covariate
Regression slopes homogeneity is verified
BSS_Pa_Crit_Mass_Ssc..y_tf ~ Itot_meso_mWm2_Ero + Mumo_txt + Cond_Fact_txt post-hoc tests
Residuals are not normal - Shapiro-Wilk: stat = 0.965, p-value = 0.0212 - (α=0.1)
Residuals variances are homogeneic - Levene: stat = 1.141, p-value = 0.34 - (α=0.1)
Residuals are homoscedastic - Harrison-McCabe: statistic = 0.519, p-value = 0.608 (α=0.1)
There are 0 outliers in **
1 Mean (SD); n

3.3 Results tables

SuppTab 3.K: Linear regressions data (ax+b equation, R² and p-value)
Respiration_rate_mWm2 Itot_meso_mWm2_Ero Biomass_gAFDWm2
All T. medium T. low T. high All T. medium T. low T. high All T. medium T. low T. high
BSS_Pa_Crit_Mass_Ssc

-1.479e-03 x +1.455
r²=0.007 (n=84)

-1.416e-02 x +1.769
r²=0.335
* (n=29)**

1.243e-03 x +1.230
r²=0.003 (n=27)

8.246e-04 x +1.483
r²=0.006 (n=28)

-2.071e-03 x +1.460
r²=0.008 (n=84)

-2.303e-02 x +1.904
r²=0.410
* (n=29)**

2.176e-03 x +1.226
r²=0.002 (n=27)

-6.459e-04 x +1.537
r²=0.002 (n=28)

-1.791e-02 x +1.557
r²=0.066 * (n=84)

-4.365e-02 x +1.796
r²=0.332
(n=29)**

4.841e-03 x +1.218
r²=0.005 (n=27)

-9.179e-03 x +1.593
r²=0.032 (n=28)

S. plana

4.411e-04 x +1.378
r²=0.000 (n=19)

-3.750e-02 x +2.223
r²=0.857
(n=6)**

1.503e-02 x +0.919
r²=0.098 (n=6)

2.145e-03 x +1.402
r²=0.025 (n=7)

-7.651e-04 x +1.417
r²=0.001 (n=19)

-3.017e-02 x +2.350
r²=0.644 · (n=6)

1.339e-02 x +0.987
r²=0.036 (n=6)

-3.265e-03 x +1.665
r²=0.034 (n=7)

-2.406e-02 x +1.702
r²=0.114 (n=19)

-5.362e-02 x +2.168
r²=0.627 · (n=6)

1.519e-02 x +1.034
r²=0.036 (n=6)

-2.432e-02 x +1.815
r²=0.166 (n=7)

S. plana & (H. diversicolor)

-5.379e-03 x +1.523
r²=0.040 (n=19)

-2.466e-02 x +1.952
r²=0.258 (n=6)

-1.155e-03 x +1.297
r²=0.002 (n=6)

-1.587e-02 x +2.075
r²=0.661 * (n=7)

-5.607e-03 x +1.504
r²=0.030 (n=19)

-3.742e-02 x +2.088
r²=0.722 * (n=6)

1.755e-02 x +1.044
r²=0.074 (n=6)

-2.028e-02 x +2.154
r²=0.647 * (n=7)

-3.982e-02 x +1.718
r²=0.203 · (n=19)

-7.047e-02 x +1.930
r²=0.698 * (n=6)

1.891e-02 x +1.120
r²=0.049 (n=6)

-6.206e-02 x +2.049
r²=0.678 * (n=7)

(S. plana) & H. diversicolor

2.063e-03 x +1.312
r²=0.026 (n=18)

-2.166e-03 x +1.512
r²=0.008 (n=6)

7.227e-03 x +1.179
r²=0.101 (n=6)

3.029e-03 x +1.198
r²=0.143 (n=6)

3.227e-03 x +1.315
r²=0.018 (n=18)

-8.963e-03 x +1.612
r²=0.040 (n=6)

2.073e-02 x +1.066
r²=0.222 (n=6)

7.630e-03 x +1.100
r²=0.154 (n=6)

1.635e-02 x +1.262
r²=0.048 (n=18)

-1.723e-02 x +1.566
r²=0.033 (n=6)

3.006e-02 x +1.148
r²=0.164 (n=6)

2.327e-02 x +1.155
r²=0.151 (n=6)

H. diversicolor

3.312e-03 x +1.188
r²=0.050 (n=17)

-4.245e-03 x +1.264
r²=0.127 (n=6)

4.482e-03 x +0.940
r²=0.069 (n=6)

-1.110e-03 x +1.812
r²=0.066 (n=5)

1.879e-02 x +0.845
r²=0.300 * (n=17)

1.922e-02 x +0.735
r²=0.231 (n=6)

7.359e-03 x +0.949
r²=0.027 (n=6)

9.326e-04 x +1.719
r²=0.012 (n=5)

2.000e-02 x +1.154
r²=0.037 (n=17)

5.236e-02 x +0.797
r²=0.286 (n=6)

1.139e-02 x +0.988
r²=0.021 (n=6)

-1.929e-04 x +1.754
r²=0.000 (n=5)

BSS_Pa_Crit_Mass_Visu

1.745e-03 x +1.262
r²=0.010 (n=82)

1.184e-03 x +1.240
r²=0.003 (n=28)

-1.004e-03 x +1.268
r²=0.002 (n=27)

9.002e-04 x +1.384
r²=0.004 (n=27)

2.567e-03 x +1.252
r²=0.011 (n=82)

-5.259e-04 x +1.277
r²=0.000 (n=28)

-7.232e-03 x +1.348
r²=0.027 (n=27)

1.948e-03 x +1.352
r²=0.009 (n=27)

-3.278e-03 x +1.338
r²=0.002 (n=82)

-1.478e-03 x +1.279
r²=0.000 (n=28)

-9.608e-03 x +1.323
r²=0.021 (n=27)

-2.152e-03 x +1.439
r²=0.001 (n=27)

S. plana

7.409e-03 x +1.097
r²=0.228 * (n=19)

1.294e-02 x +0.911
r²=0.437 (n=6)

-3.169e-04 x +1.225
r²=0.000 (n=6)

4.252e-03 x +1.318
r²=0.112 (n=7)

5.547e-03 x +1.124
r²=0.121 (n=19)

1.235e-02 x +0.809
r²=0.463 (n=6)

-1.069e-02 x +1.403
r²=0.054 (n=6)

-9.221e-04 x +1.553
r²=0.003 (n=7)

1.544e-03 x +1.292
r²=0.001 (n=19)

2.114e-02 x +0.894
r²=0.418 (n=6)

-1.267e-02 x +1.371
r²=0.059 (n=6)

-1.357e-02 x +1.683
r²=0.058 (n=7)

S. plana & (H. diversicolor)

-1.624e-02 x +1.703
r²=0.195 · (n=18)

3.943e-03 x +1.091
r²=0.015 (n=6)

-1.395e-02 x +1.506
r²=0.135 (n=6)

-3.373e-02 x +2.471
r²=0.617 · (n=6)

-1.548e-02 x +1.626
r²=0.127 (n=18)

-2.689e-02 x +1.779
r²=0.832 * (n=6)

-9.005e-03 x +1.376
r²=0.011 (n=6)

-4.110e-02 x +2.667
r²=0.526 (n=6)

-6.234e-02 x +1.825
r²=0.273 * (n=18)

-5.194e-02 x +1.678
r²=0.845
(n=6)**

-1.758e-02 x +1.402
r²=0.024 (n=6)

-1.261e-01 x +2.457
r²=0.557 · (n=6)

(S. plana) & H. diversicolor

-1.551e-05 x +1.379
r²=0.000 (n=17)

8.505e-03 x +1.131
r²=0.135 (n=6)

1.527e-03 x +1.357
r²=0.005 (n=6)

-7.415e-04 x +1.365
r²=0.006 (n=5)

5.552e-04 x +1.367
r²=0.000 (n=17)

-1.135e-03 x +1.424
r²=0.001 (n=6)

6.742e-03 x +1.299
r²=0.024 (n=6)

4.565e-03 x +1.178
r²=0.038 (n=5)

5.437e-03 x +1.337
r²=0.005 (n=17)

1.662e-03 x +1.391
r²=0.000 (n=6)

8.239e-03 x +1.336
r²=0.013 (n=6)

7.970e-03 x +1.261
r²=0.012 (n=5)

H. diversicolor

3.383e-03 x +1.165
r²=0.074 (n=18)

-2.132e-03 x +1.311
r²=0.101 (n=6)

1.074e-02 x +0.659
r²=0.597 · (n=6)

-5.749e-03 x +1.929
r²=0.403 (n=6)

1.108e-02 x +0.993
r²=0.227 * (n=18)

1.385e-02 x +0.952
r²=0.379 (n=6)

1.150e-02 x +0.791
r²=0.099 (n=6)

-6.087e-03 x +1.859
r²=0.206 (n=6)

1.311e-02 x +1.185
r²=0.025 (n=18)

3.610e-02 x +1.008
r²=0.430 (n=6)

2.048e-02 x +0.830
r²=0.102 (n=6)

-2.559e-02 x +1.841
r²=0.263 (n=6)

SuppTab 3.L: Significative differences between regression slopes
Respiration_rate_mWm2 Itot_meso_mWm2_Ero Biomass_gAFDWm2
BSS_Pa_Crit_Mass_Ssc

pairwise ~ Cond_Fact_txt

T. medium - T. high *
T. low - T. high ·

T. medium - T. high
T. low - T. high

-

pairwise ~ Cond_Fact_txt | Mumo_txt

H. diversicolor T. medium - T. high ·
H. diversicolor T. low - T. high ·

H. diversicolor T. medium - T. high ·
H. diversicolor T. low - T. high *

H. diversicolor T. low - T. high ·

4 Supplementary data

4.1 Session information

─ Session info ───────────────────────────────────────────────────────────────
 setting  value
 version  R version 4.4.0 (2024-04-24 ucrt)
 os       Windows 11 x64 (build 22631)
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 quarto   1.4.554 @ C:\\PROGRA~1\\Quarto\\bin\\quarto.exe

─ Packages ───────────────────────────────────────────────────────────────────
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